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MiR-34 regulated genes and pathways as targets for therapeutic intervention

A gene and gene expression technology, applied in the fields of molecular biology and medicine, can solve the problems of unknown regulatory pathways and network details

Inactive Publication Date: 2010-08-11
MIRNA THERAPEUTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As mentioned above, the current limitation of this approach is that many details of the regulatory pathways and networks affected by any given miRNA, including hsa-miR-34, remain unknown

Method used

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  • MiR-34 regulated genes and pathways as targets for therapeutic intervention
  • MiR-34 regulated genes and pathways as targets for therapeutic intervention
  • MiR-34 regulated genes and pathways as targets for therapeutic intervention

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Experimental program
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Effect test

preparation example Construction

[1156] B. Preparation of nucleic acid

[1157] Nucleic acids can be prepared by any technique known to those of ordinary skill in the art, such as, for example, chemical synthesis, enzymatic production, or biological production. The specific consideration is that the miRNA probe of the present invention is chemically synthesized.

[1158] In some embodiments of the invention, miRNAs are recovered or isolated from biological specimens. The miRNA may be recombinant or it may be natural or endogenous to the cell (produced from the genome of the cell). Consider that biological specimens can be processed in a certain way to improve the recovery of small RNA molecules such as miRNA. U.S. Patent Application Serial No. 10 / 667,126 describes such methods, and this application is specifically incorporated herein by reference. Generally, the method involves lysing the cells with a solution with guanidine salt and detergent.

[1159] Optionally, nucleic acid synthesis is performed according t...

Embodiment 1

[1200] Analysis of gene expression after transfection with HSA-MIR-34A

[1201] It is believed that miRNA regulates gene expression by binding to targeted mRNA transcripts, and (1) initiating degradation of the transcript or (2) altering protein translation from the transcript. The upward or downward changes in protein expression caused by translation regulation can cause changes in downstream gene products and the activity and expression of genes regulated by these proteins in turn. These numerous regulatory effects are revealed as changes in the total mRNA expression profile. Perform microarray gene expression analysis to identify genes that are misregulated by hsa-miR-34a.

[1202] For each of the three time points, synthetic precursor miR-34a (Ambion) or two negative control miRNAs (precursor miR-NC1, Ambion cat.no.AM17110 and precursor miR-NC2, Ambion, cat.no .AM17111) was reverse transfected into four samples of A549 cells. Cells were transfected with siPORT NeoFX (Ambion...

Embodiment 2

[1206] Cell pathways affected by HSA-MIR-34a

[1207] The misregulation of gene expression caused by hsa-miR-34a affects many cellular pathways (Table 1), which represent potential therapeutic targets for the control of cancer and other diseases and disorders. The inventors have determined the names and characteristics of cellular gene pathways affected by the regulatory cascade induced by hsa-miR-34a expression. Use Ingenuity pathway analysis (Version 4.0, Systems, Redwood City, CA) perform cell pathway analysis. The Fisher's exact test (Fisher, 1922) is used to determine the change of the designated pathway. The pathways most significantly affected after hsa-miR-34a overexpression in A549 cells are shown in Table 2.

[1208] These data prove that hsa-miR-34a directly or indirectly affects the expression of a large number of cancer-related, cell proliferation-related, cell development-related, cell signaling-related and cell cycle-related genes, and therefore mainly affects ca...

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Abstract

The present invention concerns methods and compositions for identifying genes or genetic pathways modulated by miR-34, using miR-34 to modulate a gene or gene pathway, using this profile in assessing the condition of a patient and / or treating the patient with an appropriate miRNA.

Description

[0001] This application claims the priority of US Provisional Patent Application No. 60 / 942,971 filed on June 8, 2007, which is incorporated herein by reference in its entirety. Technical field [0002] The invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to methods and compositions for the treatment of diseases or disorders affected by miR-34 microRNA, microRNA expression, and genes and cell pathways directly and indirectly regulated by it. Background technique [0003] In 2001, several research groups used cloning to isolate and identify a large group of "microRNA" (miRNA) from C.elegans, Drosophila and humans (Lagos-Quintana et al., 2001; Lau et al. , 2001; Lee and Ambros, 2001). Hundreds of miRNAs have been identified in plants and animals (including humans) that do not seem to have endogenous siRNA. Therefore, although similar to siRNA, miRNA is different. [0004] The length of miRNAs observed so far is appr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00G01N33/574C12N15/113
CPCC12N2320/12C12N2320/31C12N2310/141C12N15/113G01N2800/52G01N33/574C12N2330/31Y10T436/143333A61P1/00A61P11/00A61P13/00A61P17/00A61P21/00A61P25/00A61P27/02A61P31/04A61P31/10A61P31/12A61P33/00A61P35/00A61P35/02A61P37/00A61P43/00A61P5/00A61P7/00A61P9/00
Inventor 安德里斯·G·巴德卢娜·帕特拉瓦拉麦克·拜罗姆查尔斯·D·约翰逊大卫·布朗
Owner MIRNA THERAPEUTICS
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